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Papers In Press, published online ahead of print March 20, 2002
J. Biol. Chem, 10.1074/jbc.M111771200
Submitted on December 10, 2001
Revised on January 25, 2002
Accepted on March 20, 2002
transcription in hepatocytes
Department of Biochemistry and Molecular Pharmacology, West Virginia University, Morgantown, WV 26506-9142
Corresponding Author: fbhillgartner{at}hsc.wvu.edu
In previous work, we characterized a 3,5,3’-triiodothyronine response element (T3RE) in acetyl-CoA carboxylase-
(ACC) promoter 2 that mediated 3,5,3’-triiodothyronine (T3) regulation of ACC transcription in chick embryo hepatocytes. Sequence comparison analysis revealed the presence of a sterol regulatory element-1 (SRE-1) located 5 bp downstream of the ACC T3RE. Here, we investigated the role of this SRE-1 in modulating T3 regulation of ACC transcription. Transfection analyses demonstrated that the SRE-1 enhanced T3-induced ACC transcription by more than 2-fold in hepatocytes. The effect of the SRE-1 on T3 responsiveness required the presence of the T3RE in its native orientation. In pull-down experiments, the mature form of SREBP-1 specifically bound the -isoform of the nuclear T3 receptor (TR), and the presence of T3 enhanced this interaction. A region of TR containing the DNA binding domain plus flanking sequences (amino acids 21-157) was required for interaction with SREBP-1, and a region of SREBP-1 containing the basic-helix-loop-helix-leucine zipper domain (amino acids 300 to 389) was required for interaction with TR. In gel mobility shift experiments, TR, retinoid X receptor-, and mature SREBP-1 formed a tetrameric complex on a DNA probe containing the ACC T3RE and SRE-1, and the presence of T3 enhanced the formation of this complex. Formation of the tetrameric complex stabilized the binding of SREBP-1 to the SRE-1. These results indicate that SREBP-1 interacts with TR·RXR in an orientation-specific manner to enhance T3-induced ACC transcription in hepatocytes. T3 regulation of ACC transcription in nonhepatic cell cultures such as chick embryo fibroblasts is markedly reduced compared to that of chick embryo hepatocytes. Here, we also show that alterations in SREBP expression play a role in mediating cell-type-dependent differences in T3 regulation of ACC transcription.
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